| 質粒名稱: | MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 163719 |
| 存儲實驗室: | Matthew Bennett |
|
載體骨架基本信息 |
|
| 載體名稱: | CloDF13 |
| 載體出品公司: | -- |
| 空載體大小: | -- |
| 完整質粒大小: | -- |
| 載體修飾: | -- |
| 載體類型: | 細菌表達 |
| 篩選標記: | -- |
| 載體抗性: | 氯霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | araC dnaQ926.1TAG dam.1TAG seqA.1TAG emrR.1TAG ugi.1TAG CDA1.1TAG |
| 物種來源: | Synthetic |
| 插入大小: | -- |
| 基因突變: | dnaQ926.1TAG大壩。1標簽序列。1標記emrR。1塔格ugi。1TAG CDA1.1TAG；每個基因的起始蛋氨酸密碼子后插入琥珀色密碼子 |
| GenBank編號 | -- |
| 啟動子: | -- |
| 標簽/融合蛋白: | -- |
| 克隆方法: | Gibson克隆 |
| 5’克隆位點: | -- |
| 3’克隆位點: | -- |
| 5’測序引物 | -- |
| 3’測序引物 | -- |
|
文獻引用方法 |
|
| 材料和方法部分: | MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG was a gift from Matthew Bennett (Addgene plasmid # 163719 ; //n2t.net/addgene:163719 ; RRID:Addgene_163719) |
| 參考文獻部分: | A suppressor tRNA-mediated feedforward loop eliminates leaky gene expression in bacteria. Ho JML, Miller CA, Parks SE, Mattia JR, Bennett MR. Nucleic Acids Res. 2021 Mar 18;49(5):e25. doi: 10.1093/nar/gkaa1179. 10.1093/nar/gkaa1179 PubMed 33290521 |
MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG
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MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG載體質粒基本信息
MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG質粒圖譜載體圖譜和MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG載體序列質粒序列多克隆位點信息
MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG質粒載體簡介
MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG質粒序列載體序列
LOCUS MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG 6699 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Arabinose-inducible expression of six mutagenesis genes, each
encoding one amber codon after the initiator methionine.
ACCESSION .
VERSION .
KEYWORDS MP6.6TAG_AraC_Para-dnaQ926-dam-seqA-emrR-ugi-CDA1.6TAG
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6699)
TITLE A suppressor tRNA-mediated feedforward loop eliminates leaky gene
expression in bacteria.
REFERENCE 2 (bases 1 to 6699)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..6699
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(39..777)
/direction=LEFT
/label=CloDF13 ori
/note="Plasmids containing the CloDF13 (CDF) origin of
replication can be propagated in E. coli cells that contain
additional plasmids with compatible origins."
terminator 867..912
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
CDS complement(927..1805)
/codon_start=1
/gene="araC"
/product="L-arabinose regulatory protein"
/label=araC
/translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
EKVNDVAVKLS"
promoter 1832..2116
/gene="araBAD"
/label=araBAD promoter
/note="promoter of the L-arabinose operon of E. coli; the
araC regulatory gene is transcribed in the opposite
direction (Guzman et al., 1995)"
primer_bind 2039..2058
/label=pBAD Forward
/note="For vectors with E. coli araBAD promoter, forward
primer"
CDS 4936..5193
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="M*TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVH
TAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 5223..5852
/codon_start=1
/product="cytidine deaminase from the sea lamprey
Petromyzon marinus (Rogozin et al., 2007)"
/label=PmCDA1
/translation="M*TDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGE
RRACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEK
ILEWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKI
FIQSSHNQLNENRWLEKTLKRAEKRRSELSIMIQVKILHTTKSPAV"
terminator 5879..5910
/label=tonB terminator
/note="bidirectional E. coli tonB-P14 transcription
terminator"
CDS complement(5926..6585)
/codon_start=1
/gene="cat"
/product="chloramphenicol acetyltransferase"
/label=CmR
/note="confers resistance to chloramphenicol"
/translation="MEKKITGYTTVDISQWHRKEHFEAFQSVTQCTYNQTVQLDITAFL
KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
1 cactcggtcg ctacgctccg ggcgtgagac tgcggcgggc gctgcggaca catacaaagt
61 tacccacaga ttccgtggat aagcagggga ctaacatgtg aggcaaaaca gcagggccgc
121 gccggtggcg tttttccata ggctccgccc tcctgccaga gttcacataa acagacgctt
181 ttccggtgca tctgtgggag ccgtgaggct caaccatgaa tctgacagta cgggcgaaac
241 ccgacaggac ttaaagatcc ccaccgtttc cggcgggtcg ctccctcttg cgctctcctg
301 ttccgaccct gccgtttacc ggatacctgt tccgcctttc tcccttacgg gaagtgtggc
361 gctttctcat agctcacaca ctggtatctc ggctcggtgt aggtcgttcg ctccaagctg
421 ggctgtaagc aagaactccc cgttcagccc gactgctgcg ccttatccgg taactgttca
481 cttgagtcca acccggaaaa gcacggtaaa acgccactgg cagcagccat tggtaactgg
541 gagttcgcag aggatttgtt tagctaaaca cgcggttgct cttgaagtgt gcgccaaagt
601 ccggctacac tggaaggaca gatttggttg ctgtgctctg cgaaagccag ttaccacggt
661 taagcagttc cccaactgac ttaaccttcg atcaaaccac ctccccaggt ggttttttcg
721 tttacagggc aaaagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
781 tctactgaac cgctctagat ttcagtgcaa tttatctctt caaatgtagc acctgaagtc
841 agcccaggag gaagaggaca tccggtcaaa taaaacgaaa ggctcagtcg aaagactggg
901 cctttcgttt tagacttagg gaccctttat gacaacttga cggctacatc attcactttt
961 tcttcacaac cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc
1021 gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg
1081 gtggtgctca aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg
1141 ctaatcccta actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc
1201 tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa
1261 gcctcgcgta cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc
1321 cgcagtaaca attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct
1381 tgcccggcgt taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc
1441 gggcgaaaga accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag
1501 gcgcgcggac gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg
1561 tagtgatgaa tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct
1621 cgtccctgat ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc
1681 attcccagcg gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa
1741 cccgccacca gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca
1801 gccatacttt tcatactccc accattcaga gaagaaacca attgtccata ttgcatcaga
1861 cattgccgtc actgcgtctt ttactggctc ttctcgctaa cccaaccggt aaccccgctt
1921 attaaaagca ttctgtaaca aagcgggacc aaagccatga caaaaacgcg taacaaaagt
1981 gtctataatc acggcagaaa agtccacatt gattatttgc acggcgtcac actttgctat
2041 gccatagcat ttttatccat aagattagcg gatcctacct gacgcttttt atcgcaactc
2101 tctactgttt ctccataccc gtttttttgg acgcgtacaa ctcaagtctg acataaatga
2161 ccgctatgta gagcactgca attacacgcc agatcgttct cgctaccgca accaccggta
2221 tgaaccagat tggtgcgcac tatgaaggcc acaagatcat tgagattggt gccgttgaag
2281 tggtgaaccg tcgcctgacg ggcaataact tccatgttta tctcaaaccc gatcggctgg
2341 tggatccgga agcctttggc gtacatggta ttgccgatga atttttgctc gataagccca
2401 cgtttgccga agtagccgat gagttcatgg actatattcg cggcgcggag ttggtgatcc
2461 ataacgcagc gttcgatatc ggctttatgg actacgagtt ttcgttgctt aagcgcgata
2521 ttccgaagac caatactttc tgtaaggtca ccgatagcct tgcggtggcg aggaaaatgt
2581 ttcccggtaa gcgcaacagc ctcgatgcgt tatgtgctcg ctacgaaata gataacagta
2641 aacgaacgct gcacggggca ttactcgatg cccagatcct tgcggaagtt tatctggcga
2701 tgaccggtgg tcaaacgtcg atggcttttg cgatggaagg agagacacaa cagcaacaag
2761 gtgaagcaac aattcagcgc attgtacgtc aggcaagtaa gttacgcgtt gtttttgcga
2821 cagatgaaga gattgcagct catgaagccc gtctcgatct ggtgcagaag aaaggcggaa
2881 gttgcctctg gcgagcataa tttaatatca gtaaaccgga cataacccat gtagaagaaa
2941 aatcgcgctt ttttgaagtg ggcagggggc aagtatcccc tgcttgatga tattaaacgg
3001 catttgccca agggcgaatg tctggttgag ccttttgtag gtgccgggtc ggtgtttctc
3061 aacaccgact tttctcgtta tatccttgcc gatatcaata gcgacctgat cagtctctat
3121 aacattgtga agatgcgtac tgatgagtac gtacaggccg cacgcgagct gtttgttccc
3181 gaaacaaatt gcgccgaggt ttactatcag ttccgcgaag agttcaacaa aagccaggat
3241 ccgttccgtc gggcggtact gtttttatat ttgaaccgct acggttacaa cggcctgtgt
3301 cgttacaatc tgcgcggtga gtttaacgtg ccgttcggcc gctacaaaaa accctatttc
3361 ccggaagcag agttgtatca cttcgctgaa aaagcgcaga atgccttttt ctattgtgag
3421 tcttacgccg atagcatggc gcgcgcagat gatgcatccg tcgtctattg cgatccgcct
3481 tatgcaccgc tgtctgcgac cgccaacttt acggcgtatc acacaaacag ttttacgctt
3541 gaacaacaag cgcatctggc ggagatcgcc gaaggtctgg ttgagcgcca tattccagtg
3601 ctgatctcca atcacgatac gatgttaacg cgtgagtggt atcagcgcgc aaaattgcat
3661 gtcgtcaaag ttcgacgcag tataagcagc aacggcggca cacgtaaaaa ggtggacgaa
3721 ctgctggctt tgtacaaacc aggagtcgtt tcacccgcga aaaaataatt cagctaagac
3781 actgcactgg attaagatgt agaaaacgat tgaagttgat gatgaactct acagctatat
3841 tgccagccac actaagcata tcggcgagag cgcatccgac attttacggc gtatgttgaa
3901 attttccgcc gcatcacagc ctgctgctcc ggtgacgaaa gaggttcgcg ttgcgtcacc
3961 tgctatcgtc gaagcgaagc cggtcaaaac gattaaagac aaggttcgcg caatgcgtga
4021 acttctgctt tcggatgaat acgcagagca aaagcgagcg gtcaatcgct ttatgctgct
4081 gttgtctaca ctatattctc ttgacgccca ggcgtttgcc gaagcaacgg aatcgttgca
4141 cggtcgtaca cgcgtttact ttgcggcaga tgaacaaacg ctgctgaaaa atggtaatca
4201 gaccaagccg aaacatgtgc caggcacgcc gtattgggtg atcaccaaca ccaacaccgg
4261 ccgtaaatgc agcatgatcg aacacatcat gcagtcgatg caattcccgg cggaattgat
4321 tgagaaggtt tgcggaacta tctaacggct gaaattaatg aggtcatacc caaatgtagg
4381 atagttcgtt tacgcccatt gaacaaatgc taaaatttcg cgccagccgc cacgaagatt
4441 ttccttatca ggagatcctt ctgactcgtc tttgcatgca catgcaaagc aagctgctgg
4501 agaaccgcaa taaaatgctg aaggctcagg gaattaacga gacgttgttt atggcgttga
4561 ttacgctgga gtctcaggaa aaccacagta ttcagccttc tgaattaagt tgtgctcttg
4621 gatcatcccg taccaacgcg acgcgtattg ccgatgaact ggaaaaacgc ggttggatcg
4681 aacgtcgtga aagcgataac gatcgccgct gcctgcatct gcaattaacg gaaaaaggtc
4741 acgagttttt gcgcgaggtt ttaccaccgc agcataactg cctgcatcaa ctctggtccg
4801 cgctcagcac aacagaaaaa gatcagctcg agcaaatcac ccgcaaattg ctctcccgtc
4861 tcgaccagat ggaacaagac ggtgtggttc tcgaagcgat gagctaataa tacaaaaatt
4921 aggaggaatt tcaacatgta gacaaattta tctgacatca ttgaaaaaga aacaggaaaa
4981 caactagtga ttcaagaatc aattctaatg ttaccagaag aagtagagga agtaattggg
5041 aataaaccag aaagtgatat tttagttcat actgcttatg atgaaagtac agatgaaaat
5101 gtaatgctat taacttcaga tgctccagaa tataaacctt gggctttagt aattcaagac
5161 agtaatggag aaaataaaat taaaatgtta taagtcgaga ttaagtaaac cggaatctga
5221 agatgtagac cgacgcggaa tacgttcgta tccacgaaaa actggacatc tacaccttca
5281 aaaaacagtt cttcaacaac aaaaaatctg tttctcaccg ttgctacgtt ctgttcgaac
5341 tgaaacgtcg tggtgaacgt cgtgcgtgct tctggggtta cgcggttaac aaaccgcagt
5401 ctggtaccga acgtggtatc cacgcggaaa tcttctctat ccgtaaagtt gaagaatacc
5461 tgcgtgacaa cccgggtcag ttcaccatca actggtactc ttcttggtct ccgtgcgcgg
5521 actgcgcgga aaaaatcctg gaatggtaca accaggaact gcgtggtaac ggtcacaccc
5581 tgaaaatctg ggcgtgcaaa ctgtactacg aaaaaaacgc gcgtaaccag atcggtctgt
5641 ggaacctgcg tgacaacggt gttggtctga acgttatggt ttctgaacac taccagtgct
5701 gccgtaaaat cttcatccag tcttctcaca accagctgaa cgaaaaccgt tggctggaaa
5761 aaaccctgaa acgtgcggaa aaacgtcgtt ctgaactgtc tatcatgatc caggttaaaa
5821 tcctgcacac caccaaatct ccggcggttt aaacttaatt aacggcactc ctcagccaag
5881 tcaaaagcct ccggtcggag gcttttgact acatgcccat ggcgtttacg ccccgccctg
5941 ccactcatcg cagtactgtt gtaattcatt aagcattctg ccgacatgga agccatcaca
6001 aacggcatga tgaacctgaa tcgccagcgg catcagcacc ttgtcgcctt gcgtataata
6061 tttgcccata gtgaaaacgg gggcgaagaa gttgtccata ttggccacgt ttaaatcaaa
6121 actggtgaaa ctcacccagg gattggctga gacgaaaaac atattctcaa taaacccttt
6181 agggaaatag gccaggtttt caccgtaaca cgccacatct tgcgaatata tgtgtagaaa
6241 ctgccggaaa tcgtcgtggt attcactcca gagcgatgaa aacgtttcag tttgctcatg
6301 gaaaacggtg taacaagggt gaacactatc ccatatcacc agctcaccgt ctttcattgc
6361 catacggaac tccggatgag cattcatcag gcgggcaaga atgtgaataa aggccggata
6421 aaacttgtgc ttatttttct ttacggtctt taaaaaggcc gtaatatcca gctgaacggt
6481 ctggttatag gtacattgag taactgactg aaatgcctca aaatgttctt tacgatgcca
6541 ttgggatata tcaacggtgg tatatccagt gatttttttc tccattttag cttccttagc
6601 tcctgaaaat ctcgataact caaaaaatac gcccggtagt gatcttattt cattatggtg
6661 aaagttggaa cctcttacgt gccaagccaa ataggccgt
//